Production and characterisation of monoclonal antibodies against immunoglobulins of Cirrhinus mrigala (Hamilton 1822)

Mohanty, Snatashree and Makesh, M and Rajendran, K V and Suresh Babu, P P and Anand, Deepika and Kumar, Sourav and Kumar, Abhay and Baitha, Raju and Sarma, Kamal (2020) Production and characterisation of monoclonal antibodies against immunoglobulins of Cirrhinus mrigala (Hamilton 1822). Indian Journal of Fisheries, 67 (2). pp. 55-61.

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    Abstract

    Serum immunoglobulins (Ig) of mrigal Cirrhinus mrigala (Hamilton 1822) immunised with bovine serum albumin (BSA), were purified by affinity chromatography using BSA-CL agarose column. The purified mrigal Ig (m-Ig) was characterised under reducing condition by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) which revealed two bands of 85 and 26 kDa corresponding to heavy and light chain, respectively. Following fusion of splenocytes from Balb/c mice immunised with purified m-Ig with myeloma cells, three hybridomas showing reactivity with m-Ig were cloned by limiting dilution. The monoclonal antibodies (MAbs) generated by these clones were designated as 3B2-E12, 3B2-F9 and 4C3-B2 and characterised by western blotting and isotyping. Western blot analysis of the supernatant from the three clones with purified m-Ig indicated that, all the three MAbs were specific to heavy chain. Isotyping revealed that 3B2-E12 MAb was of IgG1 isotype whereas the other two MAbs were of IgG2a isotype. Cross reactivity of anti-mrigal Ig MAb (3B2-E12) was observed with serum Ig of Catla catla and Labeo rohita indicating semi-conserved nature of Ig in Indian major carps.

    Item Type: Article
    Uncontrolled Keywords: Cirrhinus mrigala, ELISA, Immunoglobulin, Monoclonal Antibodies, Western blotting
    Subjects: Fish Biotechnology > Microbiology
    Fish Biotechnology
    Divisions: CMFRI-Karwar
    Depositing User: Arun Surendran
    Date Deposited: 03 Jul 2020 10:38
    Last Modified: 04 Jul 2020 06:11
    URI: http://eprints.cmfri.org.in/id/eprint/14337

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